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Embedding vitrification ultralow-temperature storage method for bletilla striata seeds

A technology of vitrification ultra-low temperature and preservation method, applied in the field of ultra-low temperature preservation of plant germplasm resources, to achieve high germination rate, shorten dehydration time, and achieve long-term ultra-low temperature preservation

Active Publication Date: 2014-09-03
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the cryopreservation of embedment and vitrification of Baiji seeds.

Method used

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  • Embedding vitrification ultralow-temperature storage method for bletilla striata seeds

Examples

Experimental program
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Effect test

Embodiment 1

[0032]Collect intact uncracked white and mature fruits more than 150 days after pollination, first rinse with tap water, wipe the surface with 75% ethanol and place it on an ultra-clean workbench, then soak in 75% ethanol for 30 seconds, rinse with sterile water for 2-3 times, followed by immersion in 0.1% mercuric chloride for 20 minutes, and rinsed with sterile water 4-5 times. The sterilized fruit is placed in a sterile petri dish, the surface moisture is blotted dry with sterile filter paper, the fruit pod is cut longitudinally, and the powdered seeds in the fruit pod are taken for later use.

[0033] Put the powdered seeds into the sodium alginate embedding solution, and fully stir the seeds with a glass rod to evenly distribute the seeds in it. The formula of the sodium alginate embedding solution is: VW+2% sodium alginate+2.0mol L -1 Glycerin+0.4mol·L -1 Sucrose; drop the sodium alginate embedding solution containing seeds into the calcium ion embedding solution with a...

Embodiment 2-8

[0039] In the implementation steps of embodiment 2-8, the dehydration time in PVS2 solution is respectively 40min, 60min, 80min, 100min, 120min, 140min, 160min, and other steps are identical with embodiment 1.

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Abstract

The invention relates to an embedding vitrification ultralow-temperature storage method for bletilla striata seeds. The method comprises the following steps: collecting integral fruit, cleaning, wiping surfaces with ethanol, immersing in ethanol, flushing with sterile water, immersing in a mercuric chloride solution with a concentration of 0.1%, flushing, absorbing water on the fruit surfaces, splitting capsules in a longitudinal direction, and taking out powder-like seeds; pouring seeds into a sodium alginate embedding solution, stirring uniformly, dropwise putting into a calcium ion embedding solution, and putting in a room-temperature environment and forming embedded beads through solidifying; putting the embedded beads into a PVS2 solution and performing dewatering processing for 20-160 min; putting into a refrigeration tube, adding a PVS2 solution, and then putting the refrigeration tube in liquid nitrogen for refrigeration storage; when seeds are needed, taking out the refrigeration tube, unfreezing and removing the PVS2 solution, and washing; and inoculating the washed embedded beads to a solid VW medium for culture, enabling the seed to germinate, and performing domestication transplanting after seedlings grow to have the height of 6-10 cm. The seeds obtained by employing the method are high in germination rate and the cost can be saved.

Description

technical field [0001] The invention relates to a method for cryopreservation of plant germplasm resources, in particular to a method for embedding and vitrification cryopreservation of seeds of bletilla germplasm. Background technique [0002] Baiji (Bletilla striata (Thunb.) Reichb.f.), also known as Lianjicao, Ruolan, Gangen, Baigen, Baili, Baiji, and Zihui, is a perennial herbaceous plant belonging to Orchidaceae and is distributed in Asia. Northern Myanmar passes through China to Japan and the Korean Peninsula. Baiji has purple inflorescences and dense leaves. It can be cultivated as a potted ornamental plant or as a ground cover. It can also produce fresh cut flowers and has good ornamental value. The tuber of Baiji is a commonly used traditional Chinese medicine, which has the functions of astringent and hemostasis, detumescence and granulation promotion, and promotion of wound healing. Significant curative effect. In recent years, the scope of clinical application...

Claims

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Application Information

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IPC IPC(8): A01N3/00
Inventor 王彩霞田敏
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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